The major objective of this research is to investigate the biosynthesis of the asparagine-linked oligosaccharides of the gonadtropin, the glycoprotein hormones of placenta and pituitary origin. Specifically, the sheep pituitary system will be used and the focus of attention will center upon ovine luteinizing hormone, which, unlike any other glycoprotein, has N-acetylgalactosamine as an integral part of the asparagine-linked carbohydrate unit. This objective will be approached in five different ways. First, the structure of the individual asparagine-linked carbohydrates will be extensively characterized by chemical and enzymatic methods. Second, the sheep pituitary sytem will be characterized. Preliminary experiments will be designed to compare oligosaccharide biosynthesis in a pituitary cell-free system with those of hen oviduct and other systems in which a lipid-linked intermediate is involved in glycosylation. Special attention will be directed toward isolation and complete characterization of the lipid-linked intermediates. Third, the role of glucose-containing oligosaccharides as intermediates in the process of asparagine-linked oligosaccharide assembly and attachment will be considered. Fourth, the enzymes involved in the processing, recognition, and synthesis of the inner-core will be isolated and studied. In particular, the enzyme which transfers the lipid-linked oligosaccharide to protein will be the first to be isolated in the pituitary system. This will require the use of suitable nonglycosylated acceptors and appropriate oligosaccharide-dolichol phosphate compounds. Finally, experiments will be conducted which will seek to correlate the physiological effects of such agents as LH-RH, cAMP, steroids, and phosphodiesterase.